CN105037461B - Multistage aqueous two-phase separated in synchronization Gardenoside and Gardenia Yellow - Google Patents
Multistage aqueous two-phase separated in synchronization Gardenoside and Gardenia Yellow Download PDFInfo
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- CN105037461B CN105037461B CN201510396431.9A CN201510396431A CN105037461B CN 105037461 B CN105037461 B CN 105037461B CN 201510396431 A CN201510396431 A CN 201510396431A CN 105037461 B CN105037461 B CN 105037461B
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Abstract
The present invention relates to it is a kind of in high yield and the Gardenoside of purity and the extraction separation method of Gardenia Yellow.The present invention is to provide the separated in synchronization purifying that efficiently mild Two-phase system realizes Gardenoside and Gardenia Yellow, comprise the following steps：It is that raw material obtains cape jasmine crude extract with cape jasmine fruit powder, by multistage low molecule alkoxide double-aqueous phase system extraction separation, concentrate drying obtain Gardenoside and the crystallization first product of Gardenia Yellow, using obtained after recrystallization purity be more than 98.5% cape jasmine glycosidal crystalline and color value 535 Gardenia Yellow.This method is easy to operate, green high-efficient, and economy is time saving, is easy to amplify.It has not only started application of the aqueous two phase extraction technique in separated in synchronization Gardenia Yellow and Gardenoside, but also improve Gardenoside and Gardenia Yellow isolates and purifies efficiency.
Description
Technical field
The invention belongs to the technical fields of natural product active ingredient extraction, and in particular to one kind is high from cape jasmine fruit powder to be received
Rate and the method for purity extraction Gardenoside and Gardenia Yellow.
Background technology
Gardenoside is a kind of active ingredient in cape jasmine fruit, have the effects that it is anti-oxidant, anti-inflammatory, in medicine, health care, life
Analyte detection etc. extensive application.Gardenoside is iridoid, different name geniposide.It is main in food industry
Natural edible coloring agent gardenia blue and gardenia red can be made.Gardenia Yellow is a kind of mixture, wherein main ingredient includes hiding
Carthamin, crocetin and Gardenoside.Gardenia Yellow contains certain promoting blood circulation, eliminating the phlegm, analgesic, decompression in addition to it can colour
Medical functions are waited, but when the OD values of Gardenia Yellow are more than 0.4, Gardenia Yellow is readily converted into green or fades, and wherein price is held high
Expensive crocetin, may be used on medicine trade.The extraction separation on Gardenoside and Gardenia Yellow has CO at present2- SFE, chromatography
With the method for macroporous resin adsorption.But these methods not only take, and financial cost is also very high.Due to the polarity of Gardenoside
It is similar with Gardenia Yellow, it is easily eluted out together with Gardenia Yellow when absorption method separates Gardenia Yellow, so as to influence the color of Gardenia Yellow
Valency and OD values.It is all to use the ethyl alcohol of opposed polarity to macroreticular resin to be related to the continuously patent of extraction Gardenoside and Gardenia Yellow
Gradient elution is carried out, is reached the separated purpose of the two.Therefore active development high efficiency extraction separation Gardenoside and Gardenia Yellow technology,
Will have great realistic meaning and considerable market prospects.Double-aqueous phase system（ATPE）With it with easy to operate, separation
It is efficient, to characteristics such as the destructions of separate substance zero, it has been widely used in biochemistry, cell biology, biochemical industry and day
The product separation and purifying in the fields such as the extraction of right product.We are using isopropanol-salt double-aqueous phase system extraction cape jasmine fruit raw material
Production Gardenia Yellow waste liquid obtains purity and is more than 98.5% cape jasmine glycosidal crystalline, and applies for a patent（The patent No.
201410721860.4）, have been enter into the substantive examination stage at present, but the method using aqueous two-phase extraction Gardenoside reported at present
All it is not involved with the synchronous extraction of Gardenia Yellow.For this purpose, our compositions by the double-aqueous phase system to low mass molecule alcohol/inorganic salts
It is studied, the method for establishing multistage aqueous two-phase separated in synchronization purifying gardenoside and Gardenia Yellow.This method is easy to operate, green height
Effect, economy is time saving, is easy to amplify.It has not only started aqueous two phase extraction technique answering in separated in synchronization Gardenia Yellow and Gardenoside
With, while improve Gardenoside and Gardenia Yellow isolates and purifies efficiency.
The content of the invention
It is obtained in high yield and the Gardenoside of high-purity and Gardenia Yellow using aqueous two-phase the object of the present invention is to provide a kind of
Method.Cape jasmine crude extract is obtained using cape jasmine fruit powder, multistage isopropanol/salt double-aqueous phase system extraction is passed it through and separates, is dense
Contracting is dried to obtain Gardenoside and Gardenia Yellow crystallization first product, after recrystallization, obtains the Gardenoside of high-purity and Gardenia Yellow knot
It is brilliant.
A kind of method for isolating and purifying Gardenoside, it is characterised in that：Low mass molecule alcohol and the double water of salt are prepared at room temperature
Phase system and drawing diagram, according to determining suitable double-aqueous phase system into phase stability.
Described is a kind of method for isolating and purifying Gardenoside and Gardenia Yellow, it is characterised in that：It draws used in aqueous two-phase figure
Low molecule alcohol is the alkylol of the linear chain or branch chain of C1-C5.Particularly preferably ethyl alcohol, isopropanol or propyl alcohol, the salt
For alkali salt, alkali metal salts or ammonium salt.Sylvite, sodium salt and ammonium salt can be selected from.Specifically, the sylvite can be：
Potassium carbonate, dipotassium hydrogen phosphate；Sodium salt is trisodium citrate, sodium carbonate, sodium sulphate；Ammonium salt is ammonium sulfate, ammonium carbonate.
Described is a kind of method for isolating and purifying Gardenoside and Gardenia Yellow, it is characterised in that：Low molecule is prepared at room temperature
Alcohol/salt double-aqueous phase system, cape jasmine crude extract is added in the double-aqueous phase system, under the conditions of 20-50 DEG C, 300r/min items
Temperature constant magnetic stirring 30-60min under part makes Gardenoside and Gardenia Yellow be enriched in phase.60-100min is stood, through 1000-
2000rpm centrifuges 20-40min, isolates Gardenoside and the enrichment phase of Gardenia Yellow.This is added to identical double-aqueous phase system
In lower phase, under the conditions of 20-50 DEG C, temperature constant magnetic stirring 30-60min under the conditions of 300r/min, add in auxiliary agent, make Gardenoside and
Gardenia Yellow is enriched in respectively in lower phase and upper phase, isolates Gardenoside and the enrichment phase of Gardenia Yellow.Upper phase containing Gardenia Yellow,
It is concentrated and dried through vacuum rotating to obtain Gardenia Yellow crystal crude product under the conditions of -0.04-0.09MPa, 45 DEG C；Containing under Gardenoside
It is mutually extracted using 95% ethyl alcohol, is stood overnight by alcohol extract liquor repeatedly, 30min is centrifuged through 2000rpm, except desalting ,-
0.04-0.09MPa is concentrated and dried to obtain Gardenoside crystal crude product through vacuum rotating under the conditions of 35 DEG C.Weight is carried out with organic solvent
Crystallization obtains the Gardenoside of high-purity and Gardenia Yellow crystallization.
A kind of method for isolating and purifying Gardenoside and Gardenia Yellow, it is characterised in that the auxiliary agent for hydrochloric acid, sulfuric acid,
One or more of nitric acid, acetic acid, citric acid mixture.
A kind of method for isolating and purifying Gardenoside, it is characterised in that the solvent of the recrystallization concretely third
Any one in ketone, ethyl acetate or petroleum ether, the volume of the recrystallization solvent are the first quality of Gardenoside and Gardenia Yellow
4-10 times.
The present invention isolates and purifies cape jasmine crude extract using low mass molecule alcohol-salt multistage aqueous two phase extraction technique, through recrystallization
To obtain the Gardenoside of high-purity and Gardenia Yellow crystal in high yield.This method is easy to operate, green high-efficient, economical time saving, is easy to
Amplification.It has not only started application of the aqueous two phase extraction technique in separated in synchronization Gardenia Yellow and Gardenoside, improves Gardenoside
With the efficiency that isolates and purifies of Gardenia Yellow, and the technique is also applied in the isolating and purifying of other natural pigments.
The present invention separates Gardenoside and Gardenia Yellow compared with the prior art, achieves following effect：
1. the present invention has developed a kind of multistage low mass molecule alcohol-salt Two-phase system separated in synchronization purifying gardenoside and cape jasmine
Huang, in the prior art without relevant report.
Under the conditions of the present invention, purity is 95.9%, yield is 85% Gardenoside and Gardenia Yellow crystal can be obtained, is compared
Good effect is produced in the prior art.
Specific embodiment
The method of the present invention is illustrated below by specific embodiment, but the present invention is not limited thereto.
Experimental method described in following embodiments is conventional method unless otherwise specified；The reagent and material, such as
Without specified otherwise, commercially obtain.
Embodiment 1
Double-aqueous phase system 10g is prepared for 10% with ethyl alcohol mass percent by ammonium sulfate at room temperature, takes the cape jasmine after concentration thick
Extracting solution 1ml is added in the double-aqueous phase system, temperature constant magnetic stirring（20 DEG C, 300r/min）40min, by Gardenia Yellow and Cape jasmine
Sub- glycosides is enriched in phase.60min is stood, 30min is centrifuged through 2000rpm, isolates Gardenoside and the enrichment phase of Gardenia Yellow.Again will
This is added in the lower phase of identical double-aqueous phase system, at ambient temperature, temperature constant magnetic stirring（20 DEG C, 300r/min）
40min, while add in 10w.% hydrochloric acid, makes Gardenoside and Gardenia Yellow be enriched in respectively in lower phase and upper phase, isolate Gardenoside and
The enrichment phase of Gardenia Yellow, the upper phase containing Gardenia Yellow, is concentrated and dried under the conditions of -0.04-0.09MPa, 45 DEG C through vacuum rotating
Gardenia Yellow crystal crude product is obtained, then Gardenia Yellow is obtained through re-crystallizing in ethyl acetate.Lower phase containing Gardenoside is anti-using 95% ethyl alcohol
Multiple extraction, stands overnight by alcohol extraction liquid, and 30min is centrifuged through 2000rpm, filtering, and filtrate is in -0.04-0.09MPa, and 35
It is concentrated and dried through vacuum rotating to obtain Gardenoside crystal crude product under the conditions of DEG C.It is recrystallized, is obtained with the ethyl acetate of 8 times of volumes
To Gardenoside purity and yield be respectively:98.6%, 85.8%, the purity and yield of Gardenia Yellow are respectively：98.5%,
86.5%, Gardenia Yellow color value 535.
Embodiment 2
Double-aqueous phase system 10g is prepared for 20% with isopropanol mass percent by sodium carbonate at room temperature, takes the cape jasmine after concentration
Crude extract 1ml is added in the double-aqueous phase system, temperature constant magnetic stirring（20 DEG C, 300r/min）40min, by Gardenia Yellow and
Gardenoside is enriched in phase.60min is stood, 30min is centrifuged through 2000rpm, isolates Gardenoside and the enrichment phase of Gardenia Yellow.Again
This is added in the lower phase of identical double-aqueous phase system, at ambient temperature, temperature constant magnetic stirring（20 DEG C, 300r/min）
40min adds in 30w.% hydrochloric acid, Gardenoside and Gardenia Yellow is made to be enriched in respectively in lower phase and upper phase, isolates Gardenoside and cape jasmine
Yellow enrichment phase, the upper phase containing Gardenia Yellow are concentrated and dried to obtain under the conditions of -0.04-0.09MPa, 45 DEG C through vacuum rotating
Gardenia Yellow crystal crude product, then obtain Gardenia Yellow through acetone recrystallization.Lower phase containing Gardenoside is extracted repeatedly using 95% ethyl alcohol,
It is stood overnight by alcohol extraction liquid, centrifuges 30min through 2000rpm, filtering, filtrate is under the conditions of -0.04-0.09MPa, 35 DEG C
It is concentrated and dried to obtain Gardenoside crystal crude product through vacuum rotating.It is recrystallized with the acetone of 4 times of volumes, obtained Gardenoside
Purity and yield are respectively:96.8.%, 84.9%, the purity and yield of Gardenia Yellow are respectively：97.5%, 85.5%, Gardenia Yellow color value
535。
Embodiment 3
Double-aqueous phase system 10g is prepared for 30% with isopropanol mass percent by potassium carbonate at room temperature, takes the cape jasmine after concentration
Crude extract 1ml is added in the double-aqueous phase system, temperature constant magnetic stirring（20 DEG C, 300r/min）40min, by Gardenia Yellow and
Gardenoside is enriched in phase.60min is stood, 30min is centrifuged through 2000rpm, isolates Gardenoside and the enrichment phase of Gardenia Yellow.
This is added in the lower phase of identical double-aqueous phase system, at ambient temperature, temperature constant magnetic stirring（20 DEG C, 300r/min）
40min adds in 10w.% sulfuric acid, Gardenoside and Gardenia Yellow is made to be enriched in respectively in lower phase and upper phase, isolates Gardenoside and cape jasmine
Yellow enrichment phase.Upper phase containing Gardenia Yellow, is concentrated and dried to obtain under the conditions of -0.04-0.09MPa, 45 DEG C through vacuum rotating
Gardenia Yellow crystal crude product, then recrystallize to obtain Gardenia Yellow through petroleum ether.Lower phase containing Gardenoside is extracted repeatedly using 95% ethyl alcohol
It takes, is stood overnight by alcohol extraction liquid, 30min is centrifuged through 2000rpm, through vacuum under the conditions of -0.04-0.09MPa, 35 DEG C
Spin concentration is dried to obtain Gardenoside crystal crude product.It is recrystallized with the petroleum ether of 6 times of volumes, the purity of obtained Gardenoside
It is respectively with yield:97.8.%, 85.9%, the purity and yield of Gardenia Yellow are respectively：95.5%, 86.8%, Gardenia Yellow color value 535.
Claims (4)
1. a kind of method of multistage low mass molecule alcohol/salt double-aqueous phase system separated in synchronization purifying gardenoside and Gardenia Yellow, feature exist
In producing to obtain cape jasmine crude extract for raw material using cape jasmine fruit powder；Low mass molecule alcohol-salt double-aqueous phase system is prepared at room temperature, by Cape jasmine
Sub- crude extract is added in the double-aqueous phase system, under the conditions of 20-50 DEG C, temperature constant magnetic stirring 30- under the conditions of 300r/min
60min, makes Gardenoside and Gardenia Yellow be enriched in phase, stands 60-100min, and 20-40min is centrifuged through 1000-2000rpm, point
Gardenoside and the enrichment phase of Gardenia Yellow are separated out, then this is added in the lower phase of identical double-aqueous phase system, in 20-50 DEG C of condition
Under, temperature constant magnetic stirring 30-60min under the conditions of 300r/min, while auxiliary agent is added, Gardenoside and Gardenia Yellow is made to be enriched in respectively
In lower phase and upper phase, Gardenoside and the enrichment phase of Gardenia Yellow are isolated, the upper phase containing Gardenia Yellow, in -0.04-0.09MPa, 45
It is concentrated and dried through vacuum rotating to obtain Gardenia Yellow crystal crude product under the conditions of DEG C；Lower phase containing Gardenoside is anti-using 95% ethyl alcohol
Multiple extraction, then alcohol extraction liquid is stood overnight, 30min is centrifuged through 2000rpm, except desalting, in -0.04-0.09MPa, and 35 DEG C
Under the conditions of be concentrated and dried to obtain Gardenoside crystal crude product through vacuum rotating；It is recrystallized with organic solvent, obtains high-purity
Gardenoside and Gardenia Yellow crystallization；
The auxiliary agent is the mixture of one or more of hydrochloric acid, sulfuric acid, nitric acid, acetic acid, citric acid；
The low mass molecule alcohol/salt double-aqueous phase system, low mass molecule alcohol are ethyl alcohol, isopropanol or propyl alcohol, and salt is alkali salt, alkali gold
Belong to salt or ammonium salt.
2. according to the method described in claim 1, it is characterized in that：The salt is sylvite, sodium salt or ammonium salt.
3. according to the method described in claim 2, it is characterized in that：The sylvite is dipotassium hydrogen phosphate, potassium carbonate；Sodium salt is lemon
Lemon acid trisodium, sodium carbonate, sodium sulphate；Ammonium salt is ammonium sulfate, ammonium carbonate.
4. according to the method described in claim 1, it is characterized in that：The recrystallization solvent is acetone, ethyl acetate or oil
Any one in ether, addition are 4-10 times of Gardenoside and Gardenia Yellow crude product quality.
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| CN105670336B (en) * | 2016-03-10 | 2017-06-30 | 河南中大恒源生物科技股份有限公司 | A kind of technique from the cape jasmine highly purified Gardenia Yellow of fruit high efficiency extraction |
| CN107325859A (en) * | 2017-08-16 | 2017-11-07 | 杨杰东 | Antifriction antiwear type lubricating oil additive and preparation method thereof |
| CN107722078B (en) * | 2017-10-18 | 2020-07-10 | 青岛大学 | Method for extracting, separating and purifying phlorizin by n-propanol-potassium sodium tartrate aqueous two-phase system |
| CN110357935B (en) * | 2019-07-19 | 2023-02-14 | 湖北聿津生物科技有限公司 | Method for extracting geniposide and yellow pigment step by step through continuous phase change |
| CN112480194B (en) * | 2020-11-26 | 2022-05-31 | 浙江工业大学 | Method for simultaneously preparing high-purity geniposide and gardenia yellow by one-step two-aqueous-phase extraction |
| CN112724703B (en) * | 2021-03-05 | 2022-03-04 | 中国林业科学研究院林产化学工业研究所 | Method for extracting gardenia yellow pigment |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103387489A (en) * | 2013-07-11 | 2013-11-13 | 苏州大学 | Preparation method of high pure crocin and geniposide |
| CN104356184A (en) * | 2014-12-03 | 2015-02-18 | 邵阳学院 | Method for separating and purifying geniposide by using isopropanol-salt double-aqueous-phase system |
-
2015
- 2015-07-08 CN CN201510396431.9A patent/CN105037461B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103387489A (en) * | 2013-07-11 | 2013-11-13 | 苏州大学 | Preparation method of high pure crocin and geniposide |
| CN104356184A (en) * | 2014-12-03 | 2015-02-18 | 邵阳学院 | Method for separating and purifying geniposide by using isopropanol-salt double-aqueous-phase system |
Non-Patent Citations (2)
| Title |
|---|
| 利用双水相萃取栀子黄废液中的栀子苷酶法生产栀子蓝色素的研究;梁华正 等;《生物工程》;20091231;第30卷(第3期);第155-158页 * |
| 栀子中栀子苷的分离纯化工艺研究;薛绍玲;《中国优秀硕士学位论文全文数据库（电子期刊） 工程科技I辑》;20071115;第B016-313页；正文第16-17页2.3，正文第20页3.1部分，正文第41页第6.1.2部分，正文第44页第6.2.7部分 * |
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